Melatonin plays pleiotropic roles in both animals and plants. Among them, the possible role of melatonin in the innate immune response in plants was emerging recently. As an initial study, we employed Arabidopsis to see whether melatonin is involved in the defense system against a virulent bacterial pathogen Psudomonas syringae DC3000. It was obviously observed that melatonin application of 10 μm concentration onto Arabidopsis and tobacco leaves induced various pathogenesis-related (PR) genes as well as a series of defense genes activated by salicylic acid (SA) and ethylene (ET), two key factors involved in the plant defense response compared to the mock-treated Arabidopsis and tobacco leaves, respectively. The induction of these defense-related genes in the melatonin treated Arabidopsis was well matched with an increase in resistance against pathogenic bacterium by suppressing its multiplication with about 10 fold relative over the mock-treated Arabidopsis. Furthermore, melatonin induced PR genes were almost completely or partially suppressed in npr1, ein2, and mpk6 Arabidopsis mutants indicative of SA and ET dependency of melatonin in plant defense signaling. These results suggest that melatonin may play a novel defense signaling molecule in plant-pathogen interaction